A simple and robust procedure for preparing graphene-oxide cryo-EM grids.

Journal of Structural Biology
Eugene PalovcakYifan Cheng

Abstract

Graphene oxide (GO) sheets have been used successfully as a supporting substrate film in several recent cryogenic electron-microscopy (cryo-EM) studies of challenging biological macromolecules. However, difficulties in preparing GO-covered holey carbon EM grids have limited their widespread use. Here, we report a simple and robust method for covering holey carbon EM grids with GO sheets and demonstrate that these grids can be used for high-resolution single particle cryo-EM. GO substrates adhere macromolecules, allowing cryo-EM grid preparation with lower specimen concentrations and provide partial protection from the air-water interface. Additionally, the signal of the GO lattice beneath the frozen-hydrated specimen can be discerned in many motion-corrected micrographs, providing a high-resolution fiducial for evaluating beam-induced motion correction.

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Citations

Aug 20, 2020·Nature·Christian B BillesbølleAashish Manglik
May 28, 2019·Proceedings of the National Academy of Sciences of the United States of America·Katerina NaydenovaChristopher J Russo
Feb 26, 2019·The Journal of Biological Chemistry·Dmitry Lyumkis
Jan 30, 2020·Nature Communications·Liming ZhengHailin Peng
Feb 14, 2020·Nature·Kheewoong BaekBrenda A Schulman
Dec 28, 2019·Proceedings of the National Academy of Sciences of the United States of America·Yimo HanNieng Yan
Sep 12, 2020·Proceedings of the National Academy of Sciences of the United States of America·Feng WangDavid A Agard
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Mar 6, 2021·IScience·Domen KampjutLeonid A Sazanov
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Sep 28, 2021·ACS Nano·Liming ZhengHailin Peng

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